Detection of methylated cpg rich sequences diagnostic for...

C - Chemistry – Metallurgy – 07 – H

Patent

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C07H 21/00 (2006.01) C12P 19/34 (2006.01) C12Q 1/04 (2006.01) C12Q 1/25 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2435770

The present invention provides methods for determining the methylation status of CpG-containing dinucleotides on a genome-wide scale using infrequent cleaving, methylation sensitive restriction endonucleases and two-dimensional gel electrophoretic display of the resulting DNA fragments. Such methods can be used to diagnose cancer, classify tumors and provide prognoses for cancer patients. The present invention also provides isolated polynucleotides and oligonucleotides comprising CpG dinucleotides that are differentially methylated in malignant cells as compared to normal, non-malignant cells. Such polynucleotides and oligonucleotides are useful for diagnosis of cancer. The present invention also provides methods for identifying new DNA clones within a library that contain specific CpG dinucleotides that are differently methylated in cancer cells as compared to normal cells.

La présente invention concerne des procédés de détermination de l'état de méthylation de dinucléotides contenant CpG à une échelle de la largeur du génome au moyen d'endonucléases de restriction sensibles à la méthylation, à coupure rare et de l'affichage bidimensionnel de l'électrophorèse sur gel des fragments d'ADN résultants. Ces procédés peuvent être utilisés pour diagnostiquer le cancer, classer des tumeurs et pour effectuer des pronostics pour des patients atteints de cancer. La présente invention concerne également des polynucléotides et des oligonucléotides isolés qui comprennent des dinucléotides CpG qui sont méthylés de manière différente dans des cellules malignes comparativement à des cellules normales, non malignes. De tels polynucléotides et oligonucléotides sont utiles pour le diagnostic du cancer. Cette invention concerne également des procédés d'identification de nouveaux clones d'ADN dans une banque, ces nouveaux clones d'ADN renfermant des dinucléotides CpG spécifiques qui sont méthylés différemment dans des cellules cancéreuses que dans des cellules normales. .

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